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Euthanasia and Necropsy
Published in Yuehuei H. An, Richard J. Friedman, Animal Models in Orthopaedic Research, 2020
Routine examination of synovial fluid from animals should include: a total cell count, a differential count and a ratio of WBC:RBC, mucin clot quality which is a measure of the viscosity and quality of the hyaluronic acid, a total protein and an A/G ratio, specific gravity, color, viscosity and glucose concentration.40 Total cell counts from normal joints in animals vary widely from joint to joint and the range for dogs is 0-2900 cells. The microorganisms most frequently recovered from spontaneous septic arthritis in pigs are Actinobacillus suis, Staphylococcus aureus, Haemophilus parasuis, Mycoplasma hyorhinis and hyosynoviae, Corynebacterium pyogenes and Ery-sipelothrix rhusiopathiae. Spontaneous septic arthritis in rats and mice is most frequently caused by Staphylococus sp., Mycoplasma arthriditis, Corynebacterium kutscheri and Streptobacillus moniliformis.41 Spontaneous septic arthritis is rare in carnivores except for Borelia burgdorferi in dogs.40 In osteoarthritis or rheumatoid arthritis, many biological makers released can be detected by biochemical or immunochemical assays (see Chapters 6, 18, and 19).
Integrative multiomics analysis reveals host-microbe-metabolite interplays associated with the aging process in Singaporeans
Published in Gut Microbes, 2022
Liwei Chen, Tingting Zheng, Yifan Yang, Prem Prashant Chaudhary, Jean Pui Yi Teh, Bobby K. Cheon, Daniela Moses, Stephan C. Schuster, Joergen Schlundt, Jun Li, Patricia L. Conway
In the oral microbiome, we identified 13 differentially abundant species in elderly individuals compared to young individuals (FDR p < .1, prevalence > 30%) (Figure 1d), where there were no shared differentially abundant species with the gut microbiome. Of the 13 species, some species including Streptococcus milleri, Rothia mucilaginosa, Rothia dentocariosa and Desulfobulbus oralis, had greater abundances in elderly people. Conversely, 5 species, including Prevotella melaninogenica, Prevotella fusca, Prevotella sp. oral taxon 299, Fusobacterium gonidiaformans and Actinobacillus suis, showed lower abundance in elderly subjects.
A glyco-engineering approach for site-specific conjugation to Fab glycans
Published in mAbs, 2023
Maria L. Jaramillo, Traian Sulea, Yves Durocher, Mauro Acchione, Melissa J. Schur, Anna Robotham, John F. Kelly, Marie-France Goneau, Alma Robert, Yuneivy Cepero-Donates, Michel Gilbert
While testing various bacterial sialyltransferases for their ability to catalyze the addition of sialic acid to mAbs, we observed very little modification of the Fc glycans (data not shown). However, we noticed significant incorporation of sialic acid in the case of cetuximab and we hypothesized that these bacterial sialyltransferases may be selectively modifying the Fab glycans found in this antibody. We decided to focus on the α-2,3-sialyltransferase from Actinobacillus suis (open reading frame ASU2_08685) because it expressed very well as a fusion with the Escherichia coli maltose-binding protein (construct AST-03). The specificity of AST-03 for the transfer of α-2,3-linked sialic acid to terminal galactose was confirmed by NMR analysis of a synthetic sialyllactose derivative (data not shown). The ability of AST-03 to transfer sialic acid to cetuximab was first confirmed by isoelectric focussing gel analysis (Figure S1). The migration to more acidic forms was not as extensive with AST-03 as with ST6Gal1, which was consistent with additional site-specificity for AST-03. The specificity of AST-03 for the Fab glycan of cetuximab was later confirmed by mass spectrometry (MS) (data not shown and Figure S2). We used AST-03 to develop a strategy for the incorporation of N-azidoacetylneuraminic acid (Neu5NAz) specifically into the Fab glycans of mAbs (Figure 1). Once Neu5NAz is present in a glycan, a label or a drug can be easily covalently attached by click chemistry. In contrast, the mammalian sialyltransferase ST6Gal1 can modify both the Fc and the Fab glycans, which can potentially interfere with the effector functions that are dependent on the Fc glycans.